Federica Lo Sardo
Oncogenomic and Epigenetic Unit, Regina Elena Cancer Institute, via Elio Chianesi 53, 00144, Rome, Italy.
Methods Mol Biol. 2016;1480:55-65. doi: 10.1007/978-1-4939-6380-5_5.
In higher eucaryotes, not all the genome is replicated simultaneously: there are parts of the genome that replicate at the beginning of S-phase (early S-phase), others that are replicated later. In each cell, early replicating genomic regions are alternated to late-replicating regions. In general, eucaryotic genomes are organized into structural domains where genes showing the same epigenetic state replicate at the same time.Here, we will describe the protocol that we routinely used for the analysis of replication timing of specific loci in Drosophila embryonic cell lines (S2 and S3) based on BrdU labeling and FACS sorting of different S-phase fractions (early, mid, late) of asynchronous cycling cells.
在高等真核生物中，并非所有基因组都同时复制：基因组的某些部分在S期开始时（早期S期）复制，其他部分则在之后复制。在每个细胞中，早期复制的基因组区域与晚期复制的区域交替出现。一般来说，真核生物基因组被组织成结构域，其中表现出相同表观遗传状态的基因会同时复制。在此，我们将描述我们常规用于分析果蝇胚胎细胞系（S2和S3）中特定基因座复制时间的实验方案，该方案基于对异步循环细胞不同S期部分（早期、中期、晚期）进行BrdU标记和FACS分选。
